Appropriate safety procedures should always be used with this material. Laboratory safety is discussed in the following publication: Biosafety in Microbiological and Biomedical Laboratories, 5th ed. HHS Publication No. (CDC) 93-8395. U.S. Department of Health and Human Services, Centers for Disease Control and Prevention. Washington DC: U.S. Government Printing Office (2007). The entire text is available online at http://www.cdc.gov/biosafety/publications/bmbl5/index.htm.

Biosafety classification is based on U.S. Public Health Service Guidelines, it is the responsibility of the customer to ensure that their facilities comply with biosafety regulations for their own country.

Age

14 to 14.5 day gestation embryo

Applications

These cells are provided for use as feeder cells to support the growth of purified mouse and human CD34+CD38- hematopoietic stem/progenitor cells.

Storage Conditions

liquid nitrogen vapor phase

Comments

These cells have been growth-arrested by irradiation with 12,000 rads. The cells will begin to deteriorate 2 weeks after plating and may no longer support the growth of cells. We recommend that you do not keep the cells in culture for longer than 2 weeks.

It is recommended that the feeder cells be plated 24 hours before use at 5 to 6 X 106 cells/T75 in order to obtain a 100% confluent monolayer for stem cells growth. Once the feeder cells have attached, the culture medium can be changed to accommodate the cells to be supported.

Complete Growth Medium

The base medium for this cell line is ATCC-formulated Dulbecco's Modified Eagle's Medium, Catalog No. 30-2002. To make the complete growth medium, add the following components to the base medium: 2-mercaptoethanol to a final concentration of 0.05 mM; fetal bovine serum to a final concentration of 10%.

Customers located in the state of Hawaii will need to contact the Hawaii Department of Agriculture to determine if an Import Permit is required. A copy of the permit or documentation that a permit is not required must be sent to ATCC in advance of shipment.